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ToxSci Advance Access published online on June 12, 2003
Toxicological Sciences, doi:10.1093/toxsci/kfg164
Toxicological Sciences © Society of Toxicology 2003; all rights reserved
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Received April 10, 2003; accepted May 28, 2003
© 2003 Society of Toxicology

Environmental Toxicology

Motorcycle Exhaust Particulates Enhance Vasoconstriction in Organ Culture of Rat Aortas Involve Reactive Oxygen Species

Hui-Ping Tzeng 1, Rong-Sen Yang 2, Tzuu-Huei Ueng 1, Shoei-Yn Lin-Shiau 1, and Shing-Hwa Liu 1*

1 Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan
2 Department of Orthopaedics, College of Medicine, National Taiwan University, Taipei, Taiwan

* To whom correspondence should be addressed. E-mail: shliu{at}ha.mc.ntu.edu.tw.


  Abstract

The effects of motorcycle exhaust particulate on the vasoconstriction were determined using rat thoracic aortas under organ culture condition treated with organic extracts of motorcycle exhaust particulate from a two-stroke engine. Motorcycle exhaust particulate extract (MEPE) induced a concentration-dependent enhancement of vasoconstriction elicited by phenylephrine in organ culture of both intact and endothelium-denuded aortas for 18 h. Nifedipine (a L-type Ca2+ channel blocker), manganese acetate (inorganic Ca2+ channel blocker), and staurosporine (a nonselective protein kinase C inhibitor), but not the selective PKC inhibitor chelerythrine, inhibited the enhancement of vasoconstriction by MEPE. Staurosporine has also been reported as a myosin light chain kinase (MLCK) inhibitor, so we tested whether the MLCK pathway was involved in the effect of MEPE. The results showed that ML-9 (a selective MLCK inhibitor) could inhibit the enhancement of vasoconstriction by MEPE. The phosphorylation of 20-kDa myosin light chain in primary culture of rat vascular smooth muscle cells was also enhanced by MEPE. Moreover, we also examined the role of reactive oxygen species (ROS) in the stimulatory effect of MEPE on the vasoconstriction. The antioxidant N-acetylcysteine significantly inhibited the enhancement of vasoconstriction by MEPE. A time-dependent increase in ROS production by MEPE was also detected in primary culture of at vascular smooth muscle cells. These results indicate that MEPE induces a marked enhancement of vasoconstriction in aorta under organ culture condition, and imply that a ROS-Ca2+-MLCK pathway may be involved in this MEPE-induced response.

Key Words: motorcycle exhaust particulate extract, aorta, vascular smooth muscle cell, myosin light chain kinase, reactive oxygen species .


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